Development of an indirect enzyme-linked immunosorbent assay for the detection of leptospiral antibodies in dogs.
Development of an indirect enzyme-linked immunosorbent assay for the detection of leptospiral antibodies in dogs.
复制标题
开发用于检测狗中钩端螺旋体抗体的间接酶联免疫吸附测定法。
DOI：
--
复制
发表时间：
2000
期刊：
影响因子：
--
通讯作者：
R. Lallier
中科院分区：
文献类型：
--
作者：
M. Ribotta;R. Higgins;M. Gottschalk;R. Lallier
血清学在钩端螺旋体病的诊断中起着重要作用。很少有实验室拥有进行显微凝集试验(MAT)的资源、专业知识或设施。因此，需要一种可以在任何诊断实验室使用的快速和简单的血清学检测方法。本研究应用问号钩端螺旋体波莫纳株属特异性、热稳定的抗原制剂，建立了检测犬血清中钩端螺旋体抗体的酶联免疫吸附试验。该抗原制剂可与问号钩端螺旋体布拉迪斯拉发、秋季、黄斑出血、波莫纳等血清型的兔抗血清及克氏钩端螺旋体的兔抗血清反应。对波莫纳、布拉迪斯拉发、黄斑出血、秋季、Hardjo和斑疹伤寒等5个血清型，158份1：100稀释度为阴性的犬血清，其相对特异性为95.6%。该方法对21份血清滴度为~gt；或=100的犬血清对不同血清型的相对敏感性为100%。这种分析很容易标准化，技术上比MAT更具优势，并且使用的是一种可以常规大量制备的抗原制剂。结论：该方法具有较高的敏感性，可作为犬血清钩端螺旋体抗体的初筛试验，随后用MAT对阳性结果进行确认。
Serology plays an important role in the diagnosis of leptospirosis. Few laboratories have the resources, expertise, or facilities to perform the microscopic agglutination test (MAT). Thus, there is a need for a rapid and simple serological test that could be used in any diagnostic laboratory. In this study, a genus-specific, heat-stable antigenic preparation from Leptospira interrogans serovar pomona was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of leptospiral antibodies in dog sera. This antigenic preparation reacted with rabbit antisera against L. interrogans serovars bratislava, autumnalis, icterohaemorrhagiae and pomona and with rabbit antiserum against L. kirschneri serovar grippotyphosa. The ELISA showed a relative specificity of 95.6% with 158 dog sera which were negative at a dilution of 1:100 in the MAT for serovars pomona, bratislava, icterohaemorrhagiae, autumnalis, hardjo, and grippotyphosa. The relative sensitivity of this assay with 21 dog sera that revealed serovars MAT titres of > or =100 to different serovars was 100%. This assay is easily standardized, technically more advantageous than MAT, and uses an antigenic preparation that can be routinely prepared in large amounts. It was concluded that this ELISA is sufficiently sensitive test to be used as an initial screening test for the detection of leptospiral antibodies in canine sera, with subsequent confirmation of positive test results with the MAT.